Development of antibodies for determination of alkylresorcinol metabolites in human urine and elucidation of ELISA cross-reactivity.

Alkylresorcinols (ARs) are amphiphilic phenolic lipids and their two main metabolites, 3-(3,5-dihydroxyphenyl)-propanoic acid (DHPPA) and 3,5-dihydroxybenzoic acid (DHBA), can be used as biomarkers of whole grain wheat and rye intake. The aim of this study was to develop antibodies against DHBA and DHPPA for use in ELISA analysis. Good calibration curves were obtained for ELISA using alkaline phosphatase (AP) conjugates. The highest sensitivity for DHPPA was found using a reagent combination of anti-DHPPA-BSA and DHPAA-AP in a direct ELISA (IC50=1.5µmol/L), and for DHBA using a reagent combination of anti-DHBA-OV and DHBA-AP (IC50=1.3µmol/L). Calibration was conducted in the linear range (0.3-27.4µmol/L), with limit of detection (LOD) 0.1µmol/L. Intra and inter CVs was in the range of 0.7-7.2% and 5.1-11.5%, respectively, for DHPPA and 1.3-9.4% and 3.5-20%, respectively, for DHBA. Mean recovery was 104% for DHPPA and 102% for DHBA. The ELISA method developed was then used for analysis of 120 urine samples from free-living men and women that had previously been analysed by gas chromatography-mass spectrometry (GC-MS). ELISA produced several-fold higher values than GC-MS. Application of high-resolution Orbitrap mass spectrometry (HR Orbitrap MS) allowed several compounds, including novel putative AR metabolites, to be identified, synthesised and confirmed as compounds with high ELISA cross-reactivity.

Sesamin modulates gene expression without corresponding effects on fatty acids in Atlantic salmon (Salmo salar L.).

This study examined the effects of sesamin inclusion in vegetable oil-based diets fed to Atlantic salmon (Salmo salar L.). The diets used differed in n-6/n-3 fatty acid (FA) ratio (0.5 and 1) and sesamin content (high 5.8 g/kg, low 1.16 g/kg and no sesamin). The oils used in the feeds were a mixture of rapeseed, linseed and palm oil. Fish were fed for 4 months. Fatty acids and expression of hepatic genes involved in transcription, lipid uptake, desaturation, elongation and ß-oxidation were measured. No major effects on the percentage of DHA in white muscle, liver triacylglycerol and phospholipid fraction were detected. Genes involved in ß-oxidation, elongation and desaturation were affected by sesamin addition. Limited effects were seen on any of the transcription factors tested and no effect was seen on the expression of peroxisome proliferator-activated receptors (PPAR). Expression of both SREBP-1 and SREBP-2 increased with sesamin addition. It was concluded that supplementation of fish feed with a high level of sesamin had a negative effect on the growth rate and live weight and did not alter the proportions of DHA in tissues even though gene expression was affected. Thus, more studies are needed to formulate a diet that would increase the percentage of DHA in fish without negative effects on fish growth.

Reliability of fasting plasma alkylresorcinol metabolites concentrations measured 4 months apart.

Alkylresorcinols (AR) have been suggested as specific dietary biomarkers of whole-grain wheat and rye intake. AR are metabolised to 3,5-dihydroxybenzoic acid (DHBA) and 3-(3,5-dihydroxyphenyl)-1-propanoic acid (DHPPA), which have longer apparent half-lives and were recently proposed to better reflect long-term whole-grain consumption than the intact AR. The objective of this study was to analyse the reliability--expressed by the intraclass correlation coefficient (ICC)--of AR metabolite concentrations among 100 participants from the European Prospective Investigation into Cancer and Nutrition (EPIC)-Potsdam study who provided two fasting plasma samples 4 months apart. DHBA and DHPPA concentrations were not significantly different between the first and second measurement over the 4-month period (P>0.05). The ICC was 0.32 (95% confidence interval (CI)=0.13-0.49) for DHBA and 0.37 (95%CI=0.19-0.53) for DHPPA. These results suggest that AR metabolites cannot be considered to be better biomarkers of whole-grain wheat and rye intake than the intact AR in fasting plasma (ICC=0.42).

The role of developing breast cancer in alteration of serum lipid profile.

AIMS: The major aim of this study is to examine the role of alterations in lipid profile in women developing breast cancer. This study was carried out between May 2009 and December 2010. BACKGROUND: The relationship between lipids and breast cancer is undistinguished. Until now, conflicting results have been reported on the association between lipids and risk of breast cancer development in women. MATERIALS AND METHODS: Plasma lipids (i.e., total cholesterol [TC], high-density lipoprotein [HDL], low-density lipoprotein [LDL], and triglycerides [TG] were analyzed from 60 controls and 120 untreated breast cancer patients with clinical and histopathological evidence, under aseptic conditions. Venous blood was drawn from the cases and controls and estimations of lipid profile were done utilizing the standard procedures. STATISTICAL ANALYSIS USED: Independent sample t-test to compare the mean serum levels of lipid profile and TC/HDL ratio between patients and controls. RESULTS: A significant rise in serum total cholesterol, low-density lipoprotein cholesterol, and ratio of total cholesterol: high density lipoprotein cholesterol values, whereas high density lipoprotein cholesterol and very low density lipoprotein cholesterol were not affected significantly by the breast cancer. CONCLUSIONS: The developing breast cancer might be considered as one of the factors in alterations in lipid profile levels.

Influence of dietary sesamin, a bioactive compound on fatty acids and expression of some lipid regulating genes in Baltic Atlantic salmon (Salmo salar L.) juveniles.

The effects of inclusion of sesamin / episesamin in Baltic Atlantic salmon (Salmo salar L.) diets based on vegetable oils were studied. The study was designed as a dose response study with two control diets, one diet based on fish oil (FO) and one diet based on a mixture of linseed and sunflower oil (6:4 by vol.) (MO). As experimental diets three different levels of inclusion of sesamin / episesamin (hereafter named sesamin) to the MO based diet and one diet based on sesame oil and linseed oil (SesO) (1:1 by vol.) were used. The dietary oils were mirrored in the fatty acid profile of the white muscle. Sesamin significantly decreased the levels of 18:3n-3 in the white muscle phospholipid (PL) fraction of all groups fed sesamin, no significant differences were found in the triacylglycerol fraction (TAG). Slightly increased levels of docosahexaenoic acid (22:6n-3, DHA) in PL and TAG were found in some of the sesamin fed groups. Sesamin significantly affected the expression of peroxisome proliferator-activated receptor alpha, scavenger receptor type B and hormone sensitive lipase, in agreement with previous studies on rainbow trout (Oncorhynchus mykiss) and Atlantic salmon (Salmo salar L.) hepatocytes published by our group. No significant effects on toxicological response measured as ethoxyresorufin O-deethylase activity was found. The total cytochrome P450 enzymes were significantly higher in MO 0.29 and SesO group. The amount of alpha- and gamma-tocopherols in liver and the amount of gamma-tocopherol in white muscle were significantly lower in fish fed the FO diet compared to the MO diet, but no difference after inclusion of sesamin was found in this study. Increased inclusion of sesamin increased the levels of sesamin and episesamin in the liver, but did not affect the amounts in white muscle.

Determinants of plasma alkylresorcinol concentration in Danish post-menopausal women.

BACKGROUND: Alkylresorcinols (AR), a group of phenolic lipids present in the outer parts of wheat and rye grain kernels, have been suggested as biomarkers of whole-grain wheat and rye intake. In this study, we investigated potential determinants of plasma AR concentration in a free-living population. SUBJECTS/METHODS: Non-fasting samples from post-menopausal women enrolled in the Danish Diet, Cancer and Health study (n = 360) were selected. Diet was assessed using a semi-quantitative food frequency questionnaires (FFQ) and the association between food items likely to contain AR and relevant non-dietary factors were studied by analysis of covariance models. RESULTS: The median AR concentration was 78 nmol/l (interquartile range = 106.9 nmol/l). Intake of rye bread, identified as the main determinant, was associated with 87% higher plasma total AR concentration per 100 g of bread (95% confidence interval = 46-139%). About 8-12% of the total variation (depending on the AR homologue) in plasma AR concentration was explained by the selected dietary variables. At a nutrient level, total dietary fiber and cereal fiber were significantly associated with plasma total AR concentration (P = 0.05), but only ≈2% of the total plasma AR concentration was explained by the dietary fiber or cereal fiber intake. CONCLUSIONS: In the studied population, AR plasma concentration was mainly affected by rye bread intake among investigated determinants.

Reliability of fasting plasma alkylresorcinol concentrations measured 4 months apart.

BACKGROUND/OBJECTIVES: Alkylresorcinols (ARs) have been suggested as specific dietary biomarkers of whole-grain intake. Evaluation of long-term reliability in repeatedly collected samples in population-based studies is needed to assess whether plasma AR reflect long-term exposure and may be used to study exposure-disease relationships in large-scale epidemiologic studies. The objective of this study was to analyze the reliability (reproducibility) of AR concentrations in fasting plasma measured 4 months apart. SUBJECTS/METHODS: AR concentrations (C17:0, C19:0, C21:0, C23:0 and C25:0 and total ARs) were measured in fasting plasma samples from 100 participants from the European Prospective Investigation into Cancer and Nutrition (EPIC)-Potsdam Study with an interval of 4 months between blood sampling. RESULTS: Fasting plasma AR concentrations were not significantly different between the first and second measurement over the 4-month period. Geometric means of total AR concentrations were 48.8 (95% confidence interval (CI)=41.5-57.3) nmol/l at the first and 48.7 (95% CI=41.4-57.4) nmol/l at the second measurement (P=0.99). The intraclass correlation coefficients for total AR concentrations were 0.42 (95% CI=0.25-0.57) for all, 0.55 (95% CI=0.32-0.72) for women, and 0.17 (95% CI=-0.11-0.42) for men. Comparable results were obtained for the individual homologues. CONCLUSIONS: Overall, we observed moderate reliability in concentration of total AR and individual homologues over time, although reliability was weaker in men than in women. These findings should be taken into account when using AR as blood biomarkers for whole-grain intake in large-scale epidemiologic studies.

Unusual outcome of Ludwig's angina.

This is a report of 2 cases of Ludwig's angina. An Indonesian young female patient developed severe stridor after oral examination. Then she underwent tracheostomy and developed post decannulation dyspnea due to huge surgical emphysema. The second case regards an Indian young male who developed disseminated intravascular coagulation and died from hemorrhage. The objectives of our cases presentation are to avoid mouth examination of Ludwig's angina if we are not ready for performing tracheotomy and to be aware of the possible development of disseminated intravascular coagulation and post decannulation emphysema.

Sesamin increases alpha-linolenic acid conversion to docosahexaenoic acid in atlantic salmon (Salmo salar L.) hepatocytes: role of altered gene expression.

In vitro cultivated Atlantic salmon (Salmo salar L.), hepatocytes were incubated without or with a mixture of sesamin and episesamin in order to test for possible effects on lipid metabolism. Sesamin/episesamin exposure (0.05 mM, final concentration) led to increased elongation and desaturation of (14)C 18:3n-3 to docosahexaenoic acid ((14)C 22:6n-3, DHA, P < 0.01) and down regulated gene expression of Delta6 and Delta5 desaturases compared to control treatment. Sesamin/episesamin further increased the hepatocytes capacity for fatty acid beta-oxidation of (14)C 18:3n-3 (P < 0.01) to the (14)C acid soluble products, acetate, malate and oxaloacetate, in agreement with an increased gene expression of carnitine palmitoyltransferase I. Also the gene expression of cluster of differentiation 36 was upregulated and the expression of scavenger receptor type B, peroxisome proliferator-activated receptors alpha and gamma were downregulated. The amount of triacylglycerols secreted by the cells tended to be lower in the sesamin/episesamin incubated hepatocytes than the control cells. This study shows that sesamin has favourable effects on lipid metabolism leading to increased level of DHA, which may be of interest for aquaculture use.

Sesamin supplementation increases white muscle docosahexaenoic acid (DHA) levels in rainbow trout (Oncorhynchus mykiss) fed high alpha-linolenic acid (ALA) containing vegetable oil: metabolic actions.

The effects of including an equi-mixture of sesamin and episesamin in fish diets based on vegetable oils of different fatty acid composition were examined. Sesamin/episesamin (hereafter named sesamin) was included at 0.58 g/100 g diet. The oil used in the feed was either a mixture of linseed and sunflower oils (6:4, by vol) or 100% linseed oil. Addition of sesamin increased the percentages of docosahexaenoic acid (DHA) in white muscle phospholipid and triacylglycerol fraction by up to 37% but the fatty acids in red muscle and liver were not affected. The expression of the peroxisome proliferator-activated receptor PPARalpha was significantly down regulated in the liver of the fish fed sesamin and mixed oil diet (P < 0.05). Sesamin and episesamin were detected in liver and muscle tissues of the fish that had been fed sesamin. Fish fed sesamin had elevated levels of total cytochrome P450 (CYP) enzymes and EROD activity in the liver, indicating an induction of CYP1A in this tissue. Our conclusion was that supplementation of fish feed with sesamin increased the proportions of DHA in the white muscle.

The role of genetic susceptibility in head and neck squamous cell carcinoma.

Our research is an additional genetic study to uncover the molecular mechanisms involved in head and neck squamous cell carcinoma (HNSCC) pathogenesis by studying loss of heterozygosity (LOH) and microsatellite instability (MSI) in both premalignant and malignant patients and to highlight the genotype of HNSCC in Upper Egypt. Patients with HNSCC from various parts of the world may have unique genotypes and this is the first genetic study of HNSCC in Sohag 500 KM to the south of Cairo. We performed a prospective study of 41 patients with precancerous and 79 patients with cancerous laryngeal, esophageal, nasopharyngeal, nasal and oral lesions, and 50 controls (The control patients were cases admitted for ear surgery or simple nasal surgery, from whom we took biopsy from mucosal lining of nasopharynx). The present study included 170 individuals who were admitted to the Ear, Nose and Throat department, Sohag University Hospital, Sohag, in Egypt in the period between April 2001 and March 2003. Samples which were taken by punch biopsy were frozen and stored at -80 degrees C and were subjected to histopathological examination. We investigated LOH and MSI by using six microsatellite markers located at chromosomes 3, 5, 9, and 17. The markers used were D3S1286, D9S171, D9S753, D17S654, D17S695, and CFS1-R. LOH was in all premalignant and malignant lesions at 5q33.3-q34 and 13% of Controls. LOH at 17p21 was absent in all premalignant lesions and was found in 53% of malignant lesions and 12.4% of Controls. In premalignant lesions, LOH was at 3pter-3p24.2 (73% of cases), at 9p21 (46%), at 9q21.1-22.3 (37%), and at 17p13 (37%). These percents increased in malignant lesions to 87, 80, 67, and 63%, respectively. They were 14, 19.4, 17, and 19% in controls. Examination of LOH could improve diagnosis, adds additional confidence, in HNSCC by DNA extraction from suspicious lesions in high-risk groups (smokers and alcoholics) and LOH at 3p/9p seems to be of particular value for early detection and definition of progression risk. If there are high percent of LOH at these chromosomes, active intervention should be done (chemoprevention and regular follow up head and neck examination for very early detection and management).

Phenolic glucosides in bread containing flaxseed.

Three phenolic glucosides, secoisolariciresinol diglucoside, p-coumaric acid glucoside and ferulic acid glucoside, were analyzed in commercial breads containing flaxseed. The total phenolic glucoside content ranged from 15 to 157mg/100g dry bread. Secoisolariciresinol diglucoside was the dominating phenolic glucoside with an average relative content of 62%, followed by p-coumaric acid glucoside (20%) and ferulic acid glucoside (18%). Strong positive correlations between the phenolic glucosides were found, indicating no major effect of raw material or bread-making process on the relative content of the phenolic glucosides in flaxseed.

Assessment of nasal obstruction with flexible nasal endoscopy.

OBJECTIVE: To report the value of nasal endoscopy as an outpatient procedure in the diagnosis of posterior nasal obstruction. METHODS: Over one year period, from March 2002 to March 2003, we evaluated 130 adult patients that attended the Ear, Nose and Throat Department of Sohag University Hospital in Egypt with persistent nasal obstruction via anterior rhinoscopy and flexible nasopharyngoscopy. We reported the cause and site of obstruction in relation to the choanae. We confirmed the diagnosis by CT scanning, rigid endoscopic examination under general anesthesia, and histopathological analysis of biopsies taken. RESULTS: Forty-six percent of our cases had posterior nasal obstruction, 43.5% due to post-choanal lesions (mainly adenoid), 33% due to pre-choanal lesions (mainly choanal polyps), and 23.5% due to choanal lesions (mainly choanal adenoid). CONCLUSION: We conclude that flexible nasal endoscopy is superior to visual examination in the evaluation of nasal obstruction; hence, we recommend its routine use.

Plasma antioxidant capacity among middle-aged men: the contribution of uric acid.

OBJECTIVE: Although assays of plasma antioxidant capacity encompass interactions between various antioxidants, uric acid concentration can exert a predominant effect on results. Therefore, individual differences in uric acid concentration may explain a many of the differences in antioxidant capacity. The objective of this study was to measure the antioxidant capacity of plasma samples with and without uric acid in order to provide more information about how the concept of antioxidant capacity could be applied. MATERIAL AND METHODS: Antioxidant capacity was measured using an enhanced chemiluminescence assay, and uric acid was removed from the samples using uricase. RESULTS: Antioxidant capacity was positively correlated with uric acid concentration, body mass index, waist circumference, abdominal sagittal diameter and the concentrations of insulin and triglycerides. These correlations were not evident when uric acid was eliminated from the sample, but antioxidant capacity was correlated with lipid concentration; this may partly reflect tocopherols that are transported by lipid molecules. CONCLUSIONS: The significance of the contribution of uric acid to the antioxidant capacity could differ according to the type of study. Antioxidant capacity measurements in cross-sectional studies may be presented both with and without the contribution of uric acid, because the absence of such data complicates interpretation of results when different populations are compared.

The role of multifrequency tympanometry in otitis media.

OBJECTIVE: To evaluate the diagnostic value of multifrequency tympanometry in otitis media with effusion and adhesive otitis media. METHODS: We selected 50 patients with long standing or recurrent attacks of otitis media with effusion from the outpatient clinic of the Ears, Nose and Throat (ENT) Department, Sohag Medical School, Egypt between May 2002 and December 2002. A control group was also selected, consisting of 25 patients with normal hearing levels and with no history of ENT problems. We conducted full audiological investigations in the form of pure tone audiometry, speech audiometry and immittancemetry. Immittancemetry included low probe tone frequency and multifrequency tympanometry. Each subject in the study group had undergone myringotomy and examination under microscope to decide if the case had either otitis media with effusion or adhesive otitis media. RESULTS: Resonant frequency proved to have the best performance in reflecting middle ear pathology. It was lowest in otitis media with effusion with a mean value of 428 +/- 159 Hz and it was highest in adhesive otitis media with a mean of 1336 +/- 230 Hz. CONCLUSION: Multifrequency tympanometry has an efficacy of 100% in the diagnosis of otitis media with effusion and 70% in the diagnosis of adhesive otitis media.

Frontal sinus cementifying ossifying fibroma.

Cementifying ossifying fibroma is a fibro-osseous lesion which produces cementum. Cementifying ossifying fibroma is a mesodermal, non-odontogenic tumor of ectopic multipotential periodontal membrane blast cells. Although, it is a benign bone tumor, it has an aggressive destructive behavior in patients under 20 years of age with high tendency for recurrence. Cementifying ossifying fibroma can be recognized microscopically by the presence of cementicles. This is a case of frontal sinus cementifying ossifying fibroma presenting with proptosis in a 39-year-old male.

An oligomer from flaxseed composed of secoisolariciresinoldiglucoside and 3-hydroxy-3-methyl glutaric acid residues.

A straight-chain oligomeric structure composed of five secoisolariciresinoldiglucoside (SDG) residues interconnected by four 3-hydroxy-3-methyl glutaric acid (HMGA) residues (molecular weight ca. 4000 Da) was assigned to the main lignan of flaxseed on the basis of nuclear magnetic resonance spectroscopy (NMR).

Modeling of alpha-tocopherol loss and oxidation products formed during thermoxidation in triolein and tripalmitin mixtures.

The degradation of alpha-tocopherol and the formation of alpha-tocopherol and triacylglycerol oxidation products at high temperatures (150-250 degrees C) over a heating period (0-4 h) for a model system ranging between triolein and tripalmitin were modeled by use of an experimental design. The oxidation products of alpha-tocopherol formed under these conditions were alpha-tocopherolquinone (1 .4-7.7%) and epoxy-alpha-tocopherolquinones (4.3-34.8%). The results indicate a very high susceptibility of alpha-tocopherol to capture peroxyl radicals upon oxidation, leading to the formation of polar tocopherol oxidation products. Both alpha-tocopherolquinone and epoxy-alpha-tocopherolquinones were not stable upon prolonged heating and were further degraded to other unknown oxidation products. The kinetics of alpha-tocopherol oxidation were significantly influenced by the triolein/tripalmitin ratio. By increasing the level of triacylglycerol unsaturation the rate of alpha-tocopherol recovery after heating increased significantly from 2.2 to 44.2% whereas in the meantime triacylglycerol polymerization increased from 0 to 3.7%.

Corn and sesame oils increase serum gamma-tocopherol concentrations in healthy Swedish women.

We studied the effects of dietary intervention with three vegetable oils (Linola, corn or sesame oil, all good sources of gamma-tocopherol) on absolute and relative concentrations of alpha- and gamma-tocopherol in human serum. The oils contained only small amounts of linolenic acid but varying amounts of oleic and linoleic acids, and they had different concentrations of alpha-tocopherol. Forty healthy female students (mean age 26 y) were randomly assigned to one of three groups and consumed a diet that contained one of the three oils for 4 wk. Refined oils were distributed as ingredients in specially prepared buns, in margarine or as dressing. Serum tocopherols, serum lipoproteins and plasma malondialdehyde concentrations were measured. The gamma-tocopherol concentrations normalized to serum lipids increased significantly in the corn and sesame oil groups (P < 0.01), and the alpha-/gamma-tocopherol ratios decreased significantly from baseline concentrations in all groups (P < 0.05). The alpha-tocopherol concentrations did not change during the diet period in any of the three groups. Serum cholesterol, serum apolipoprotein B and plasma malondialdehyde concentrations decreased significantly only in the Linola oil group (P < 0.05). These data show that a moderately modified natural diet that contains both alpha- and gamma-tocopherol increases the serum gamma-tocopherol concentration in healthy women without affecting the serum alpha-tocopherol concentration.

Phenolic compounds in berries of black, red, green, and white currants (Ribes sp.).

Multiple health benefits associated with phenolic compounds have raised the interest in the contents of these plant metabolites in foods. Several phenolic compounds were quantified from berries of Ribes nigrum (black and green currants) and Ribes x pallidum (red and white currants), by using sequential extraction with ethyl acetate and methanol and an optimized reversed-phase HPLC method with diode array detection. The highest contents of anthocyanins (3,011 mg/kg fresh weight, expressed as the aglycon) and flavonol glycosides (100 mg/kg) were found in black currant. The lack of anthocyanins in the colorless (green, white) berries was associated with increased levels of phenolic acids, especially p-coumaric acid (80 mg/kg in green currant vs. 45 mg/kg in black currant) and 4-hydroxybenzoic acid (18 mg/kg in white currant vs. 3 mg/kg in red currant). Previously, proanthocyanidins have not been quantified from berries. This study showed that the contents of extractable (22-41 mg/kg) and nonextractable proanthocyanidins (32-108 mg/kg) are comparable to those of other phenolics, with the exception of anthocyanins in black currant. Our results suggest that anthocyanins dominate in black and red currants, whereas proanthocyanidins and phenolic acids are the predominant phenolic compounds in green and white currants.